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Biomedical subjects

D J Finnegan

Publications and source records attributed to D J Finnegan.

10 recordsLinked to original sources

Transposable elements.

Transposable elements comprise a major fraction of eukaryotic genomes. They are studied both because of their intrinsic biological interest and because they can be exploited as valuable research tools. Many interesting papers dealing with various aspects of the biology of these elements have been published during the past year and a number of new elements have been reported. Four areas in which particularly valuable contributions have been made are the mechanisms of transposition, the regulation of transposition, the use of transposable elements as research tools, and the biological function of transposable elements.

Animals

Polypeptide components of Drosophila small nuclear ribonucleoprotein particles.

In eukaryotes splicing of pre-mRNAs is mediated by the spliceosome, a dynamic complex of small nuclear ribonucleoprotein particles (snRNPs) that associate transiently during spliceosome assembly and the splicing reaction. We have purified snRNPs from nuclear extracts of Drosophila cells by affinity chromatography with an antibody specific for the trimethylguanosine (m3G) cap structure of snRNAs U1-U5. The polypeptide components of Drosophila snRNPs have been characterized and shown to consist of a number of proteins shared by all the snRNPs, and some proteins which appear to be specific to individual snRNP particles. On the basis of their apparent molecular weight and antigenicity many of these common and particle specific Drosophila snRNP proteins are remarkably conserved between Drosophila and human spliceosomes. By probing western blots of the Drosophila snRNP polypeptides with a number of antisera raised against human snRNP proteins, Drosophila polypeptides equivalent to many of the HeLa snRNP-common proteins have been identified, as well as candidates for a number of U1, U2 and U5-specific proteins.

Animals

Evidence for retrotransposition of the I factor, a LINE element of Drosophila melanogaster.

LINEs are transposable elements found in various eukaryotes such as plants, protists, insects, and mammals. Their transposition is usually difficult to study, particularly in humans, where some diseases have been shown to result from LINE insertion mutations. This is due to the fact that most copies of any particular family of elements are defective and that their transposition frequency is low. By contrast, the I factor of Drosophila melanogaster transposes at high frequency during I-R hybrid dysgenesis and is a good model for studying the LINE element superfamily. LINEs encode putative polypeptides showing similarities with viral reverse transcriptases but, unlike viral retrotransposons, they do not have terminal repeats and their ability to transpose by reverse transcription has previously only been inferred from structural analysis. Here we present direct evidence for LINE retrotransposition. Transposition of an I factor marked by an intron resulted in accurate removal of the intron.

Animals

Identification of a potential RNA intermediate for transposition of the LINE-like element I factor in Drosophila melanogaster.

The I factor, a transposable element related to mammalian LINEs, controls the I-R system of hybrid dysgenesis in Drosophila melanogaster. It transposes at high frequency in the germ-line of the female progeny of crosses between females of the reactive class of strains and males of the inducer class. The structure and DNA sequence of the I factor suggest that it transposes by reverse transcription of an RNA intermediate. Northern blot and S1 mapping experiments show that a full-length RNA of the I factor is synthesized specifically in the conditions of which I factors transpose. This RNA has all characteristics of a transposition intermediate. It is only found in the ovaries of dysgenic females suggesting that I factor activity is restricted to this tissue because of regulation at the level of the initiation of transcription or RNA stability.

Animals

Eukaryotic transposable elements and genome evolution.

The changes in DNA sequence that have taken place during the evolution of eukaryotic genomes cannot be accounted for simply by base substitutions; some more complex mutations must have taken place as well. Transposable elements can affect gene structure and expression in several ways that suggest that they may have contributed to these evolutionary events.

Base Sequence

Transposable elements controlling I-R hybrid dysgenesis in D. melanogaster are similar to mammalian LINEs.

I-R hybrid dysgenesis in D. melanogaster is controlled by transposable elements known as I factors. We have determined the base sequences of one complete I factor and the ends of six others. The ends of these elements are highly conserved and are flanked by target site duplications varying in length from 10-14 bp. There are no terminal repeats, and the 3' end of one strand is A-rich, having 4-7 tandem repeats of the sequence TAA. This sequence organization is similar to that of mammalian LINEs, or L1 elements. The complete I factor sequence contains two long open reading frames, ORF1 and ORF2, of 1278 and 3258 bp. ORF1 encodes a possible nucleic acid-binding protein, and part of the amino acid sequence of ORF2 is similar to that of viral reverse transcriptases and polypeptides encoded by L1 elements. These results suggest that I factors transpose by reverse transcription of a full-length RNA.

Amino Acid Sequence

Characterization of six cloned DNAs from Drosophila melanogaster, including one that contains the genes for rRNA.

pDm plasmids were constructed from D. melanogaster and pSC101 DNAs by a modification of the EcoR1-ligase method which insured that each hybrid molecule contained a single segment of D. melanogaster chromosomal DNA (Dm segment). The sequences in the Dm segments of six cloned pDm DNAs were mapped within the D. melanogaster polytene chromosomes by in situ hybridization, and their repetition frequencies within the Dm segment and within the genome were determined. Four of these segments consist of sequences that are confined to single chromomeric regions in the polytene chromosomes and exhibit little or no repetition. The characteristics of this group, and also two of three Dm segments analyzed earlier (Wensink et al., 1974), are inconsistent with tandem repetition models of the chromomere. By contrast, the other two Dm segments contain moderately repetitive sequences that are located in the heterochromatin. One of these appears to be a segment of the Y chromosome in which about half the sequences are nonrepetitive and half are repeated about 33 times per genome, though they are not repeated within the segment. The second contains the DNA coding for 18 and 28S rRNA.

Animals